Green S J, Tarone G, Underhill C B
Department of Anatomy and Cell Biology, Georgetown University Medical Center, Washington, D.C. 20007.
Exp Cell Res. 1988 Oct;178(2):224-32. doi: 10.1016/0014-4827(88)90393-x.
To examine the role of the hyaluronate receptor in cell to cell adhesion, we have employed the K-3 monoclonal antibody (MAb) which specifically binds to the hyaluronate receptor and blocks its ability to interact with hyaluronate. In the first set of experiments, we investigated the spontaneous aggregation of SV-3T3 cells, which involves two distinct mechanisms, one of which is dependent upon the presence of divalent cation and the other is independent. The divalent cation-independent aggregation was found to be completely inhibited by both intact and Fab fragments of the K-3 MAb. In contrast, the K-3 MAb had no effect on the divalent cation-dependent aggregation of cells. In a second set of experiments, we examined alveolar macrophages. The presence of hyaluronate receptors on alveolar macrophages was demonstrated by the fact that detergent extracts of these cells could bind [3H]hyaluronate, and this binding was blocked by the K-3 MAb. Immunoblot analysis of alveolar macrophages showed that the hyaluronate receptor had a Mr of 99,500, which is considerably larger than the 85,000 Mr for that on BHK cells. When hyaluronate was added to suspensions of alveolar macrophages, the cells were induced to aggregate. This effect was inhibited by the K-3 MAb, suggesting that the hyaluronate-induced aggregation was mediated by the receptor.
为了研究透明质酸受体在细胞间黏附中的作用,我们使用了K-3单克隆抗体(MAb),它能特异性结合透明质酸受体并阻断其与透明质酸相互作用的能力。在第一组实验中,我们研究了SV-3T3细胞的自发聚集,这涉及两种不同的机制,其中一种依赖于二价阳离子的存在,另一种则与之无关。发现K-3单克隆抗体的完整形式和Fab片段均可完全抑制不依赖二价阳离子的聚集。相反,K-3单克隆抗体对细胞依赖二价阳离子的聚集没有影响。在第二组实验中,我们检测了肺泡巨噬细胞。肺泡巨噬细胞上存在透明质酸受体这一点可通过以下事实证明:这些细胞的去污剂提取物能够结合[3H]透明质酸,并且这种结合被K-3单克隆抗体阻断。对肺泡巨噬细胞的免疫印迹分析表明,透明质酸受体的分子量为99,500,这比BHK细胞上的透明质酸受体分子量85,000大得多。当向肺泡巨噬细胞悬液中添加透明质酸时,细胞会被诱导聚集。这种效应被K-3单克隆抗体抑制,表明透明质酸诱导的聚集是由该受体介导的。